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Biological Control of Weevils (Strophosoma Melangrammum and S. capitatum) in Greenery Plantations in Denmark

Contents

Preface

Dansk sammendrag

Summary

1 Introduction

• 1.1 Background
• 1.2 Project objectives
  • 1.2.1 Specific project objectives

2 Virulence of hyphomycete fungi against Strophosoma melanogrammum and S.capitatum

• 2.1 Definitions and terms used in laboratory bioassays with entomopathogenic fungi
• 2.2 Bioassay methodology
• 2.3 Analysis of bioassay data
  • 2.3.1 Concentration response bioassays (LC₅₀)
  • 2.3.2 Correlated observations over time at one concentration
• 2.4 Bioassays with adult Strophosoma melanogrammum
  • 2.4.1 Determination of average survival time at different temperatures
  • 2.4.2 Determination of lethal concentration of M. anisopliae BIPESCO 5
• 2.5 Bioassays with Strophosoma spp. larvae
  • 2.5.1 Determination of lethal time
• 2.6 Discussion
• 2.7 Conclusions

3 Implementation of molecular methods for identification of Strophosoma spp. larvae

• 3.1 Introduction
  • 3.1.1 The molecular methods
  • 3.1.2 Analysis of molecular sequence data
• 3.2 Materials and methods
  • 3.2.1 Extraction and amplification of DNA
  • 3.2.2 Search in Genbank and related bases with BLAST
  • 3.2.3 Alignments BLAST, T-Coffee and ClustalW
  • 3.2.4 Design of Primers for the cytochrome oxidase subunits I and II
• 3.3 Results
  • 3.3.1 DNA extraction from weevils
  • 3.3.2 Weevil DNA sequences and PCR amplification
  • 3.3.3 Search in Genbank and related bases with BLAST
  • 3.3.4 Alignments BLAST, T-Coffee and ClustalW
• 3.4 Discussion
• 3.5 Conclusion

4 Biology of Strophosoma melanogrammum and S. capitatum

• 4.1 Feeding periods of adult weevils
  • 4.1.1 Materials and methods
  • 4.1.2 Results
• 4.2 Larval drop from the canopy
  • 4.2.1 Materials and methods
  • 4.2.2 Results
• 4.3 Dispersal of Strophosoma adults and larvae
  • 4.3.1 Materials and methods
  • 4.3.2 Results
• 4.4 Development time of S. melanogrammum and S. capitatum
  • 4.4.1 Materials and methods
  • 4.4.2 Results
• 4.5 Discussion
  • 4.5.1 Life cycle
• 4.6 Conclusions

5 Effects of hyphomycete fungi against Strophosoma spp. in greenery plantations

• 5.1 Soil application of Metarhizium anisopliae against larvae
  • 5.1.1 Materials and methods
  • 5.1.2 Results
• 5.2 Soil application of Metarhizium anisopliae against ovipositing females
  • 5.2.1 Materials and methods
  • 5.2.2 Results
• 5.3 Soil application of B. bassiana and of M. anisopliae against adults emerging from the soil in spring
  • 5.3.1 Materials and methods
  • 5.3.2 Results
• 5.4 Application of tree trunks with lanes of Beauveria bassiana
• 5.5 Discussion
  • 5.5.1 Control of larvae
  • 5.5.2 Control of adult weevils
  • 5.5.3 Prospects for development of a microbial control strategy
• 5.6 Conclusions

6 Effects of fungi on non-target invertebrates and spatio-temporal distribution

• 6.1 Non-target effects
  • 6.1.1 Materials and methods
  • 6.1.2 Results
• 6.2 Spatio-temporal distribution
  • 6.2.1 Materials and methods
  • 6.2.2 Results
• 6.3 Discussion
• 6.4 Conclusions

7 Discussion and Conclusions

• 7.1 Population studies
• 7.2 Molecular characterization
• 7.3 Bio-assays and strain selection
• 7.4 Field releases and biocontrol experiments
• 7.5 Non-target effects
• 7.6 Conclusions

8 Perspectives

• 8.1 Biological control in Danish greenery and Christmas tree plantations
• 8.2 The status of insect pathogenic fungi in biological control
• 8.3 The grower's situation
• 8.4 Occupational health effects
• 8.5 Consumers' preferences
• 8.6 The final societal evaluation

9 References

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Version 1.0 November 2004, © Danish Environmental Protection Agency